Applying the indicator amino acid oxidation technique in the domestic cat: results of a pilot study and development of a non-steady state prediction model.

The aim of this study was to evaluate whether the indicator amino acid oxidation (IAAO) method could be applied in the domestic cat. Six adult male cats were used in a replicated 3×3 Latin square design. Three semi-synthetic diets were developed: a methionine (Met) and total sulfur AA (TSAA) deficient diet (T-BASAL; 0.24% Met+Cys - DM basis) and two Met and TSAA-sufficient diets in which either DL-Met (T-DLM) or 2-hydroxy-4-(methylthio)-butanoic acid (T-MHA) were supplemented, respectively, on an equimolar basis to meet the TSAA requirement (0.34%; NRC, 2006). After a 2-d diet adaptation, IAAO studies were performed. Cats were offered 13 small meals. The 6 th meal contained a priming dose (4.8 mg/kg-BW) of L-[1- 13C]-Phe and the remaining meals a constant dose (1.04 mg/kg-BW). Breath samples were collected every 25 min to measure 13CO2 enrichment. The following morning, fasted blood samples were collected. Cats returned to the T-BASAL top dressed with a DL-Met solution for 4d prior to being fed a new dietary treatment. Isotopic steady state was evaluated through visual inspection. Data were analyzed using PROC GLIMMIX procedure in SAS 9.4. While 13CO2 enrichment was successfully captured in breath samples, cats failed to reach 13CO2 steady state. Thus, a non-steady state isotope model was developed and coded in ACSLX (V3.1.4.2) individually for each cat on each study day to predict 13CO2 enrichment, and then, calculate oxidation of L-[1- 13C]-Phe (F 13CO2). A higher predicted F 13CO2 was observed for cats fed T-BASAL compared to the others (P<0.05), while no differences were observed between T-DLM and T-MHA (P>0.05). Cats fed T-DLM tended to have higher plasma Met concentrations compared to those fed T-BASAL with cats fed T-MHA intermediate (P=0.0867). Plasma homocysteine concentrations were higher in cats fed T-BASAL compared to the others (P<0.05), while threonine concentrations tended to be higher in cats fed T-BASAL compared to those fed T-MHA (P=0.0750). In conclusion, short-term provision of a semi-synthetic diet deficient in Met may elicit a metabolic response aiming to conserve Met. The successful quantification of 13CO2 enrichment in breath and the higher predicted F 13CO2 in cats fed a Met deficient diet suggest that the IAAO technique may be used in cats. Adaptations in the isotope protocol should be made to achieve 13CO2 steady state in breath and avoid mathematical modeling to predict F 13CO2.